KR20020044194A - Inhibition agent of tyrosinase composed of houttuynia cordata extract and cosmetic composition having whitening effect containing the same - Google Patents
Inhibition agent of tyrosinase composed of houttuynia cordata extract and cosmetic composition having whitening effect containing the same Download PDFInfo
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Abstract
Description
본 발명은 어성초 추출물로 이루어진 티로시나제 저해제 및 이를 함유하는 미백화장료에 대한 것이며, 또한 이의 제법에 관한 것이다.The present invention relates to a tyrosinase inhibitor composed of Echochocho extract, and to a whitening cosmetic containing the same, and also to a preparation method thereof.
일반적으로 사람의 피부색은 피부의 두께, 멜라닌, 카로틴 및 헤모글로빈의 양에 따라 결정되어 지는데 이중 멜라닌이 가장 결정적인 요소이다.In general, the skin color of a person is determined by the thickness of the skin, the amount of melanin, carotene and hemoglobin, of which melanin is the most decisive factor.
멜라닌은 표피내 기저층에 존재하는 색소세포인 멜라닌 세포(Melanocyte)에서 합성되며 주변 각질 세포(Keratinocyte)로 전이되어 사람의 피부색을 나타낸다.Melanin is synthesized in melanocytes (Melanocytes), which are pigment cells present in the basal layer of the epidermis, and are transferred to peripheral keratinocytes (Keratinocytes) to represent human skin color.
멜라닌 세포는 신경능에서 기원한 세포로서 멜라닌을 생성하여 피부를 자외선으로부터 보호하는 중요한 기능을 수행하고 있다. 멜라닌세포는 수지상 돌기를 갖는 등 신경세포와 형태학적으로 유사하며 많은 신호전달물질, 성장인자등에 대한 수용체를 공통적으로 갖고 있어 발생학적 기원이 신경세포와 같음을 보여주는 많은 특징들이 있다.Melanocytes are cells of neural capacity that produce melanin and play an important role in protecting the skin from UV rays. Melanocytes are morphologically similar to neurons, such as dendritic processes, and have many characteristics that show that their developmental origin is the same as neurons because they have a common receptor for many signaling agents and growth factors.
멜라닌 세포는 자외선, 염증등의 외부조건, 호르몬등 여러 가지 인자의 영향을 받고 있다. 또한, 멜라닌 세포로부터 멜라닌이 생성되기 위해서는 세포내의 티로신을 기질로 하여 티로시나제(Tyrosinase)라는 효소가 작용하여 도파퀴논(DOP Aquinone)을 생성시키며, 도파퀴논으로부터 자발적인 반응과 효소반응을 거쳐 공중합체인 흑색색소 멜라닌이 생성된다.Melanocytes are affected by various factors such as external conditions such as ultraviolet rays, inflammation, and hormones. In addition, in order to produce melanin from melanocytes, an enzyme called tyrosinase acts as a substrate to generate dopaquinone (DOP Aquinone) by using the intracellular tyrosine as a substrate. Melanin is produced.
멜라닌이 비정상적으로 적게 생산되면 백반증(Vitiligo)과 같은 피부병변이 유발되며, 반대로 과잉 생산되면 기미, 주근깨, 색소침착등과 같은 피부색소 이상 침착 증상이 발생된다.When abnormally low melanin is produced, skin lesions such as vitiligo are caused. On the contrary, excessive production of skin pigment abnormalities such as blemishes, freckles, and pigmentation occurs.
자외선에의 노출등에 의해 발생된 과도한 멜라닌 색소침착을 감소시켜 주기 위해서, 종래로부터 아스코르빈산, 코직산, 알부틴, 히드로퀴논등과 같은 티로시나제 저해활성을 가진 물질들이 화장료나 의약품에 배합사용하여 왔다. 그러나, 아스코르빈산은 비교적 높은 티로시나제 활성저해 효과를 갖지만, 분자자체의 안정성이 낮아 미백원료로 적합하지 않고, 코직산은 티로시나제 활성부위에 포함되어 있는 구리이온에 킬레이트되어 효소활동을 저해하는 작용을 하는데 성능이 우수한 반면 빛 또는 산소등에 의해 유발되는 산화반응으로부터 안정성에 문제가 많아 화장료로서 적당하지 않다.In order to reduce excessive melanin pigmentation caused by exposure to ultraviolet rays, substances having tyrosinase inhibitory activities such as ascorbic acid, kojic acid, arbutin, hydroquinone and the like have been conventionally used in cosmetics and pharmaceuticals. However, ascorbic acid has a relatively high inhibitory effect on tyrosinase activity, but is not suitable as a whitening material due to its low molecular stability, and kojic acid is chelated with copper ions contained in the tyrosinase active site to inhibit enzyme activity. While excellent in performance, it is not suitable as a cosmetic because of problems in stability from oxidation reactions caused by light or oxygen.
또한, 히드로퀴논은 기미, 주근깨, 반점 및 임신기 색소침착과잉(hyperpigmentation)과 같은 과잉색소증 치료에 국부적으로 사용되고 있으나 강력한 멜라닌 생성저해활성을 보이는 반면 색소세포의 변성 또는 치사를 일으키고 세포 본래의 기능을 손상시키는 등의 부작용이 나타나 화장품에서는 사용이 금지되고 있다.Hydroquinone is also used topically for the treatment of hyperpigmentation such as blemishes, freckles, spots and hyperpigmentation during pregnancy, but exhibits potent melanogenesis inhibitory activity while causing denaturation or lethality of pigment cells and impairing cell original functions. Side effects such as letting out appear in cosmetics is prohibited.
이와 같이, 티로시나제 저해활성을 가진 물질들이 화장료나 의약품에 배합 사용되었으나, 피부에 대한 안전성문제, 화장료 또는 의약품에 배합시 제형 및 안정성 측면 및 미백효과의 불충분 등의 요인으로 인해 그 사용이 제한되고 있다.As such, substances having tyrosinase inhibitory activity have been used in cosmetics or pharmaceuticals, but their use is limited due to factors such as safety issues on the skin, formulation and stability aspects when formulated in cosmetics or pharmaceuticals, and insufficient whitening effects. .
이들 종래의 미백효능 물질들 외에 천연물 특히 식물 중에서 미백활성 성분을 찾기 위한 연구를 계속해왔으며, 그중 감초(일본공개특허 소 60-214721호, 소 63-23809호0, 소 64-63506호, 평 1-149706호), 계피(일본공개특허 소 63-30403호), 상백피(일본공개특허 소 55-44375호, 소 64-26507호, 소 64-83009호, 평 1-256587호), 작약(일본공개특허 소 61-24619호)등 다수의 식물추출물이 티로시나제에 작용하여 멜라닌 생성을 억제한다는 사실이 밝혀졌으나, 이들 역시 안정성, 안전성, 변색 가능성 등의 측면에서 화장품이나 의약품에 유효농도이상으로 사용 하는데는 많은 문제점을 가지고 있는 실정이다.In addition to these conventional whitening agents, research has been conducted to find whitening active ingredients in natural products, especially plants. Among them, licorice (Japanese Patent Laid-Open No. 60-214721, No. 63-23809 0, No. 64-63506, Hep. 1) -149706), Cinnamon (Japanese Patent Publication No. 63-30403), Sangbaekpi (Japanese Patent Publication No. 55-44375, Small 64-26507, Small 64-83009, Flat 1-256587), Peony (Japan Although many plant extracts have been found to act on tyrosinase to inhibit melanin production, they are also used in cosmetics and pharmaceuticals above effective concentrations in terms of stability, safety, and discoloration potential. Has many problems.
한편, 어성초는 삼백초과에 속하는 다년생 초본으로서 학명은 Houttuynia Cordata 이고, 중약대사전(상해과학기술출판사)에 따르면 약모밀, 십약 등의 다른 이름이 있다.On the other hand, Eoseongcho is a perennial herb belonging to more than three hundred, the scientific name is Houttuynia Cordata, and according to the Chinese herbal medicine dictionary (Shanghai Science and Technology Publishing House), there are other names such as medicinal wheat and ten.
삼백초과(Saururaceae)는 한국, 일본, 중국 및 북미, 멕시코 등지에 분포하고, 하기와 같이 5속 7종으로 나누어지고 있다[Wu.C. Y., Acta Phytotaxon, Sinica, 6, 222 (1957)].Three hundred (Saururaceae) is distributed in Korea, Japan, China, North America, Mexico, etc., and is divided into five genera and seven species [Wu.C. Y., Acta Phytotaxon, Sinica, 6, 222 (1957)].
Fam. Saururaceae Genus Anemopsis … A. CalifornicaFam. Saururaceae Genus Anemopsis… A. Californica
Genus Saururus … S. CernuusGenus Saururus… S. Cernuus
S. ChinensisS. Chinensis
Genus Houttuynia…H. Cordata Genus Houttuynia … H. Cordata
Genus Gymnotheca … G. chinensisGenus Gymnotheca… G. chinensis
G. involucrataG. involucrata
Genus circaeocarpus … C. SaururoidesGenus circaeocarpus… C. Saururoides
이중 한국에서 자생하고 있는 것은 삼백초(Saururus chinensis Baill)와 어성초(약모밀, Houttuynia cardata Thunb)이다. 이들은 모두 습지에서 자라는 다년초로서 삼백초는 제주도 협재, 어성초(약모밀)은 울릉도, 제주도 및 중부지방에 분포하고 있다[이창복: 대한식물도감 p252(1979)]. 현재는 이들의 인공재배가 가능해져 부산, 천안 등지에서 인공으로 재배되고 있다.Among them, three native plants (Saururus chinensis Baill) and Eoseongcho (Hakwool, Houttuynia cardata Thunb) are found in Korea. These are all perennial plants that grow in the wetlands, and three hundred seconds are distributed in Hyeopjae, Jeju Island, and Eoseongcho (Yakmomil) are distributed in Ulleungdo, Jeju Island, and the central region. At present, their artificial cultivation is possible, and they are grown artificially in Busan and Cheonan.
어성초는 잎, 줄기, 뿌리 전체에 유효성분이 함유되어 있으며 특유의 독특한 냄새를 포함한 생리활성물질들이 신진대사를 도와 혈액을 맑게 하고 신장기능을 촉진시키며, 염증 증상과 체내 독소를 배설하는 등의 기능이 있어, 이미 3000여 년 전의 옛날부터 중국, 일본 등에서 민간약으로 널리 이용되어 왔다.Eoseongcho contains active ingredients throughout the leaves, stems, and roots, and bioactive substances, including their unique odors, help metabolism to clear blood, promote kidney function, and excrete inflammatory symptoms and toxins in the body. It has been widely used as a folk medicine in China and Japan since ancient times more than 3000 years ago.
한방에서 어성초는 고혈압, 변비증, 당뇨병, 해열, 종기, 해독, 관절염, 염증성 질환과 피부질환에 이용되고 있다. 주요성분으로는 전초에 정유성분인 데카노일아세트 알데히드, 메론, 미르센 등이 있는데 이중 "데카노일아세트알데히드"는 "설파민" 항생제의 40,000배의 역가를 지니고 있어 어성초를 항생제라고 해도 과언이 아니다.Herbal herbs are used in high blood pressure, constipation, diabetes, fever, boil, detoxification, arthritis, inflammatory diseases and skin diseases. The main ingredients are essential oils such as decanoyl acetaldehyde, melon and myrsen. Among them, "decanoyl acetaldehyde" has a titer of 40,000 times that of "sulfamine" antibiotics. .
또한, 어성초 뿌리에는 쿠에르치트린, 이소린, 하이퍼린 등의 프라보노이드 물질이 있으며, 특히 비타민 C가 다량으로 함유되어 있음이 확인되었다(김근영, 전남대, DM573.96-김262 ㅇ).In addition, the roots of Eochochocho were found to contain pravonoids such as querchithrin, isoline, and hyperrin, and in particular, a large amount of vitamin C was found (Kim Geun-young, Jeonnam University, DM573.96-Kim 262).
어성초에 관한 국내학위논문으로는 어성초의 항산화작용 및 활성성분(김주정, 상명여자대학교, DM518.33-김832ㅇ)와 어성초의 화학적 조성과 항미생물활성(김근영, 전남대, DM573.96-김262ㅇ) 및 어성초추출물의 항알레르기 효과에 관한 연구(노병주, 원광대, DP519.83-노724ㅇ)가 있다.Domestic thesis regarding Eochochochosis includes antioxidant activities and active ingredients of Eochochocho (Kim, Ju-Jung, Sangmyung Women's University, DM518.33-Kim832O) and chemical composition and antimicrobial activity of Echochocho (Kim Geun-Young, Jeonnam University, DM573.96-K262 ㅇ) and studies on antiallergic effects of fish extract (No. Byung-ju, Wonkwang-dae, DP519.83-no724).
또한, 국내특허출원으로는 경구용 미용제 조성물(공개특허 10-1999-066571) 및 기미,주근깨 제거용 한방 팩 재료(공개특허 10-2000-049466)가 있는데, 이들에서는 어성초를 복합 미용 재료를 구성하는 여러가지 한방 재료 중 하나로 사용하고 있으며, 어성초의 단독 사용 또는 이에 따른 작용에 대한 기재는 없다.In addition, domestic patent applications include oral cosmetic composition (Patent No. 10-1999-066571) and herbal pack material for removing blemishes and freckles (Patent No. 10-2000-049466). It is used as one of the various herbal materials constituting, there is no description of the use of fishery vinegar alone or its action.
본 발명자는 종래의 미백효능 물질이나 티로시나제 저해활성을 가진 식물추출물 등을 화장품에 사용하는데 따르는 상기와 같은 문제점을 감안하여, 보다 우수한 티로시나제 저해활성 및 미백 효능을 갖는 물질을 찾기 위해 부단히 연구하였다. 특히, 안정성 문제를 해결하기 위하여, 오래 전부터 한방 또는 민간요법에 사용되어 안전성이 입증되어 있는 천연식물을 대상으로 미백효능 유효물질을 검색한 결과, 어성초 추출물이 기존에 알려진 효과 외에 매우 뛰어난 티로시나제 활성저해효과를 나타냄을 알아내어 본 발명을 완성하였다.The present inventors have studied steadily in order to find a substance having a superior tyrosinase inhibitory activity and a whitening effect in view of the above problems caused by using a conventional whitening active substance or a plant extract having a tyrosinase inhibitory activity in cosmetics. In particular, in order to solve the stability problem, as a result of searching for a whitening active substance against natural plants that have been used for herbal or folk medicine for a long time and have proved its safety, the extract of Echochocho extract is very effective in inhibiting tyrosinase activity. The present invention was completed by finding an effect.
도 1은 실시예 2의 어성초 추출물과 알부틴의 티로시나제 활성저해 효과를 나타낸 그래프이고,1 is a graph showing the tyrosinase inhibitory effect of Echochocho extract and Arbutin of Example 2,
도 2는 어성초 추출물의 멜라닌 생합성 정도를 스트렙토마이세스 비키니엔시스(Streptomyces bikiniensis) 균주를 사용하여 실험한 결과를 사진으로 나타낸 것이다.2 is a photograph showing the results of experiments using the Streptomyces bikiniensis strain of melanin biosynthesis of Eochocho extract.
따라서, 본 발명에 의하면, 어성초 추출물로 이루어진 티로시나제 저해제 및 이를 함유하는 미백화장료가 제공된다.Therefore, according to the present invention, there is provided a tyrosinase inhibitor and a whitening cosmetic containing the same.
본 발명에서 어성초 추출물의 재료로는 어성초의 뿌리, 줄기, 잎을 모두 사용할 수 있으며, 뿌리를 사용하는 것이 효과 면에서 가장 바람직하다.In the present invention, as the material of Eochocho extract, all roots, stems, and leaves of Echochocho can be used, and it is most preferable to use the root in terms of the effect.
어성초 추출 용매로서는 물, 에탄올, 메탄올, 아세톤, 프로판올, 1,3-부틸렌글리콜을 단독 또는 혼합 사용할 수 있으며, 어성초 원료 kg당 10 내지 20ℓ를 가하고, 35℃ 내지 45℃에서 8시간 추출하는 것이 바람직하다. 이 때, 냉각콘덴서가 달린 추출기 등에서 추출을 실시할 수 있다.Water, ethanol, methanol, acetone, propanol, and 1,3-butylene glycol can be used alone or as a mixed solvent, and 10-20 L per kg of raw fish paste is added, followed by extraction at 35 ° C to 45 ° C for 8 hours. desirable. At this time, extraction can be performed in an extractor equipped with a cooling capacitor.
추출이 완료되면 1차 여과하고, 실온(20 내지 25℃)에서 7일간 방치하여 숙성시킨다. 이러한 숙성을 통하여 유효성분의 용출이 용이해지므로, 숙성을 거치지 않은 경우보다 더욱 우수한 미백 효과를 얻을 수 있게 되는데, 7일이 초과되면 한약 냄새가 없어지나 유효성분 함량이 더 이상 증가되지 않으며, 7일 미만이면 한약 냄새가 많이 남아있을 뿐 아니라 유효 성분 함량도 낮아 바람직하지 못하다.After the extraction is completed, primary filtration is carried out, and aged at room temperature (20-25 ° C.) for 7 days. This aging facilitates the dissolution of the active ingredient, so that a better whitening effect can be obtained than without the aging process. After 7 days, the odor of herbal medicine is lost, but the active ingredient content is no longer increased, and 7 days Chinese medicine less than Not only is there a lot of odor left, but the active ingredient content is low, which is undesirable.
숙성된 추출물은 다시 여과 후 농축한다. 농축은 45 내지 50℃의 온도, 25 내지 35cmHg의 감압조건에서 실시하는 것이 바람직한데, 이 범위에서 농축하고자 하는 유효성분이 다량 배출되기 때문이다. 또한, 농축은 용매가 완전히 제거된 점조성 있는 암갈색 또는 흑갈색 액체가 될 때까지 실시한다.Aged extract is again filtered and concentrated. Concentration is preferably carried out at a temperature of 45 to 50 ℃, a reduced pressure of 25 to 35cmHg, because a large amount of the active ingredient to be concentrated in this range is discharged. Concentration is also carried out until the solvent is a viscous dark brown or dark brown liquid with complete removal of the solvent.
본 발명에 의한 어성초 추출물은 우수한 티로시나제 저해제로서, 미백효과가 뛰어나다. 따라서, 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스 팩 등 미백 관련 화장료 조성물에 모두 첨가할 수 있으며, 그 양은 화장료의 건조중량에 대하여 0.0005 ∼ 20 중량% 정도로 한다. 20 중량% 초과 배합시에는 미백 효과 증가 및 안정성 면에서 경제적이지 못하며, 0.0005 중량% 미만으로 배합시에는 미백 효과가 거의 없다.Eochocho extract according to the present invention is an excellent tyrosinase inhibitor, and has an excellent whitening effect. Therefore, it can be added to all whitening-related cosmetic compositions, such as a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence pack, and the quantity is made into about 0.0005-20 weight% with respect to the dry weight of a cosmetic. When it is more than 20% by weight, it is not economical in terms of increased whitening effect and stability, and when it is blended at less than 0.0005% by weight, there is little effect of whitening.
이하, 실시예에 의해 본 발명을 보다 상세히 설명한다. 이들 실시예는 본 발명의 예시적인 기재일 뿐이며 본 발명의 범위가 이들 실시예에 국한되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are merely illustrative of the present invention and the scope of the present invention is not limited to these examples.
실시예Example
본 발명에 사용된 원료로는, 부산에서 인공재배 되어 자연건조시킨 어성초를 사용하였고, 뿌리를 동일 중량으로 사용해 화장품 원료로 추출하였다.As a raw material used in the present invention, the fish was artificially cultivated in Busan and dried naturally, and the root was used as the same weight and extracted as a cosmetic raw material.
실시예 1Example 1
음건세절(陰乾細切)한 어성초 뿌리 100g를 냉각콘덴서가 달린 추출기에서 물 2L로 40℃, 8시간 추출한 후, 400메쉬 표준체로 여과하고 실온(20 내지 25℃)에서 7일간 방치하여 숙성시킨 후, 다시 400메쉬 표준체를 이용하여 여과하였다. 이 추출물을 감압증발기로 감압농축하여 점조성이 있는 어성초 뿌리추출물 15g을 얻었다.After extracting 100g of dried dried Echochocho roots with 40 liters of water and 8 hours in an extractor equipped with a cooling condenser, it was filtered through a 400 mesh standard and left at room temperature (20 to 25 ℃) for 7 days to mature. It was again filtered using a 400 mesh standard. The extract was concentrated under reduced pressure with a reduced pressure evaporator to obtain 15 g of Echo root root extract with viscous consistency.
실시예 2Example 2
음건세절(陰乾細切)한 어성초뿌리 100g를 수욕상에서 메탄올 2L로 40℃에서 8시간 추출한 후, 400메쉬 표준체로 여과하고 실온에서 7일간 방치하여 숙성시킨 후, 다시 400메쉬 표준체를 이용하여 여과하였다. 이 추출물을 감압증발기로 감압농축하여 점조성이 있는 어성초 뿌리추출물 18.7g을 얻었다.100 g of dried edible roots were dried for 8 hours at 40 ° C. with 2 L of methanol in a water bath, filtered through a 400 mesh standard, left to mature for 7 days at room temperature, and filtered again using a 400 mesh standard. . The extract was concentrated under reduced pressure with a reduced pressure evaporator to obtain 18.7 g of root extract of viscous vinegar.
실시예 3Example 3
음건세절(陰乾細切)한 어성초뿌리 100g를 냉각콘덴서가 달린 추출기에서 물 1L, 1,3-부틸렌글리콜 1L을 이용하여 40℃에서 8시간 추출한 후, 400메쉬 표준체로 여과하고 실온에서 7일간 방치하여 숙성시킨 후, 다시 400메쉬 표준체를 이용하여 여과하였다. 이 추출물을 감압증발기로 감압농축하여 점조성이 있는 어성초뿌리추출물 20.6g을 얻었다.100 g of dried edible roots were dried for 8 hours at 40 ° C. using 1 L of water and 1 L of 1,3-butylene glycol in an extractor equipped with a cooling condenser, filtered through a 400 mesh standard, and 7 days at room temperature. After standing to mature, it was filtered again using a 400 mesh standard. The extract was concentrated under reduced pressure with a reduced pressure evaporator to obtain 20.6 g of viscous vinegar extract with viscous consistency.
실시예 4Example 4
음건세절(陰乾細切)한 어성초뿌리 100g을 수욕상에서 에탄올 2L로 40℃에서 8시간 추출한 후, 400메쉬 표준체로 여과하고 실온에서 7일간 방치하여 숙성 시킨 후, 다시 400메쉬 표준체를 이용하여 여과하였다. 이 추출물을 감압증발기로 감압농축하여 점조성이 있는 어성초뿌리추출물 19.4g을 얻었다.100 g of dried edible roots were dried for 8 hours at 40 ° C. with 2 L of ethanol in a water bath, filtered through a 400 mesh standard, left to mature at room temperature for 7 days, and filtered again using a 400 mesh standard. . The extract was concentrated under reduced pressure with a reduced pressure evaporator to obtain 19.4 g of a sticky root extract with viscous consistency.
실시예 5Example 5
음건세절(陰乾細切)한 어성초뿌리 100g을 수욕상에서 메탄올 800㎖, 아세톤 200㎖, 프로판올 200㎖, 에탄올 800㎖을 이용하여 40℃에서 8시간 추출한 후, 400메쉬 표준체로 여과하고 실온에서 7일간 방치하여 숙성시킨 후, 다시 400메쉬 표준체를 이용하여 여과하였다. 이 추출물을 감압증발기로 감압농축하여 점조성이 있는 어성초뿌리추출물 19.1g을 얻었다.100 g of dried edible roots were dried for 8 hours at 40 ° C. using 800 ml of methanol, 200 ml of acetone, 200 ml of propanol, and 800 ml of ethanol in a water bath, filtered through a 400 mesh standard, and filtered at room temperature for 7 days. After standing to mature, it was filtered again using a 400 mesh standard. The extract was concentrated under reduced pressure with a reduced pressure evaporator to obtain 19.1 g of a sticky root extract with viscous consistency.
실시예 6Example 6
음건세절(陰乾細切)한 어성초 줄기, 잎 100g을 수욕상에서 에탄올 2ℓ로 40℃에서 8시간 추출한 후, 400메쉬 표준체로 여과하고 실온에서 7일간 방치하여 숙성시킨 후, 다시 400메쉬 표준체를 이용하여 여과하였다. 이 추출물을 감압증발기로 감압농축하여 점조성이 있는 어성초 줄기, 잎 추출물 18.4g을 얻었다.100 g of dried stems and leaves of dried echinacea were extracted with 2 liters of ethanol in a water bath at 40 ° C. for 8 hours, filtered through a 400 mesh standard, left to stand at room temperature for 7 days, and then aged again using a 400 mesh standard. Filtered. The extract was concentrated under reduced pressure using a vacuum evaporator to obtain 18.4 g of a sticky stem and leaf extract with viscous consistency.
제조예 1(유연화장수)Preparation Example 1 (Flexible Cosmetic)
실시예 1에서 얻은 어성초 추출물을 함유하는 유연화장수(스킨로션)를 다음과 같이 제조하였다.Softening water (skin lotion) containing the Echo extract obtained in Example 1 was prepared as follows.
상기 조성에 따라 수상(상기원료 1-4)과 에탄올상(상기원료 5-7)을 구분하여 계량한 후, 따로 용해하고 에탄올상을 수상에 넣으며 혼합하였다.According to the composition, the aqueous phase (the raw material 1-4) and the ethanol phase (the raw material 5-7) were separated and weighed, and then separately dissolved and mixed with the ethanol phase in the aqueous phase.
이어서 첨가제(상기원료 8-9)를 넣고 혼합하였다. 이를 멤브레인 필터를 사용하여 여과한 후 적당한 용기에 담아 제품화 하였다.Subsequently, an additive (raw material 8-9) was added and mixed. This was filtered using a membrane filter and put into a suitable container to produce a product.
제조예 2 (영양화장수)Preparation Example 2 (Nutrition Cosmetics)
실시예 1에서 얻은 어성초추출물을 함유하는 영양화장수(밀크로션)를 다음과 같이 제조하였다.Nutritious longevity (wheat lotion) containing the fish extract obtained in Example 1 was prepared as follows.
상기조성에 따라 수상(상기원료 1-6)과 유상(상기원료 7-11)을 구분하여 계 량한 후, 80±2℃에서 따로 혼합용해하고 유상을 수상에 넣으며 혼합하였다.According to the composition, water phase (raw material 1-6) and oil phase (raw material 7-11) were weighed separately, and then mixed and dissolved separately at 80 ± 2 ° C., and the oil phase was mixed with water.
이어서 중화제(상기원료 12)투입하여 중화시킨 후 50℃까지 냉각시켰다. 냉각 후 상기원료 13을 투입한 다음 35℃까지 냉각시켜 적당한 용기에 담아 제품화 하였다.Subsequently, a neutralizing agent (the above raw material 12) was charged and neutralized, and then cooled to 50 ° C. After cooling, the raw material 13 was added and then cooled to 35 ° C. in a suitable container to produce a product.
제조예 3(영양크림)Preparation Example 3 (Nutrition Cream)
실시예 1에서 얻은 어성초추출물을 함유하는 영양크림을 하기 성분을 사용하여 제조예 2와 같은 방법으로 제조하였다.Nourishing cream containing the fish extract obtained in Example 1 was prepared in the same manner as in Preparation Example 2 using the following ingredients.
제조예 4(에센스)Preparation Example 4 (Essence)
실시예 1에서 얻은 어성초추출물을 함유하는 에센스를 다음과 같이 제조하였다.Essence containing the fish extract obtained in Example 1 was prepared as follows.
상기조성에 따라 원료를 차례대로 게량한 후 혼합용해하고, 이를 200메쉬체로 여과한 후 적당한 용기에 담아 제품화하였다According to the above composition, the raw materials were weighed one by one, mixed and dissolved, filtered through 200 mesh sieves, and put into a suitable container for commercialization.
시험예 1Test Example 1
상기 실시예 1∼5에서 얻은 어성초추출물에 대한 티로시나제 활성저해 효과와 알부틴의 티로시나제 활성저해 효과를 측정 비교하였다.The tyrosinase inhibitory effect and arbutin tyrosinase inhibitory effect on fish herb extracts obtained in Examples 1 to 5 were measured and compared.
효소티로시나제는 버섯류(Mushroom)에서 분리 및 정제된 것으로시그마(Sigma)사의 것을 사용하였다. 먼저 기질인 티로신(sigma제)을 증류수에 0.3㎎/㎖의 용액으로 만들고, 이 용액을 1.0㎖씩 취해 시험관에 넣었다. 여기에 인산칼륨완충액(potassium phosphate buffer : pH 6.8, 0.1M) 1.0㎖와 각 추출물의 단계별 희석용액 0.7㎖씩을 반응용액에 넣은 뒤 37℃ 항온기에서 10분간 반응시켰다. 희석용매는 에탄올과 물을 3:7의 비율로 사용하며, 대조군으로는 각 추출물 대신 희석용매만을 0.7㎖ 넣은 것을 사용하였다.Enzyme tyrosinase was isolated and purified from Mushroom (Mushroom) was used Sigma (Sigma). First, the substrate tyrosine (manufactured by sigma) was made into a solution of 0.3 mg / ml in distilled water, and 1.0 ml of this solution was taken in a test tube. Here, 1.0 ml of potassium phosphate buffer (potassium phosphate buffer: pH 6.8, 0.1M) and 0.7 ml of the dilution solution of each extract were added to the reaction solution, followed by reaction for 10 minutes at 37 ° C. in a thermostat. Diluted solvent was used in a ratio of 3: 7 ethanol and water, as a control was used to put only 0.7ml diluent solvent instead of each extract.
이 반응액에 티로시나제(1,250 unit/㎖) 0.1㎖씩을 넣고 다시 37℃ 항온기에서 10분간 반응시켰다. 이 반응액이 들어있는 시험관을 얼음물 속에 넣어서 급냉시켜 반응을 중지시키고, 흡광광도계(160-A, shimazu제)로 파장 475㎚에서 흡광도를 측정하였다.0.1 ml each of tyrosinase (1,250 unit / ml) was added to the reaction solution, and the resultant was reacted for 10 minutes at 37 ° C. in a thermostat. The test tube containing the reaction solution was placed in iced water, quenched to stop the reaction, and the absorbance was measured at a wavelength of 475 nm with an absorbance spectrometer (160-A, manufactured by shimazu).
각 추출물의 티로시나제 활성저해효과는 하기의 공식으로 구하였다.The tyrosinase inhibitory effect of each extract was determined by the following formula.
상기의 공식에 의해 구해진 각 시료의 단계별 희석용액의 티로시나제 저해율로부터 티로시나제 활성을 50% 감소시키는 각 시료의 용량 (IC50= Inhibition Concentration 50%)을 구하였다.The dose of each sample (IC 50 = Inhibition Concentration 50%) that reduced tyrosinase activity by 50% was determined from the tyrosinase inhibition rate of the dilution solution of each sample obtained by the above formula.
시험결과는 하기 표1에 나타낸 바와 같다.The test results are shown in Table 1 below.
상기 표에서 나타난 바와 같이, 어성초추출물은 알부틴에 비해 매우 강력한 미백효과를 나타내었다.As shown in the table, Echochocho extract showed a very strong whitening effect compared to arbutin.
시험예 2Test Example 2
상기 실시예 1에서 얻은 어성초추출물과 알부틴 및 코직산에 대한 B16멜라노마 세포의 멜라닌 생합성 저해시험을 하였다. 마우스 유래 B16멜라노마 세포(American Type Culture Collection, 기탁번호:6323)를 포도당 4.5g/L, 10%소태아혈청 1% 항생제가 함유된 DMEM(Dulbecco's Modified Eagles Medium(Gibco, USA))배지에 접종하여 50㎖ T-플라스크에 37℃에서 배양하였다. 5% CO2조건하에서 24시간 배양한 후, 0.02% EDTA가 함유된 0.05%트립신을 처리하여 세포를 분리하고, 이어서 50㎖ T-플라스크에 접종하여 48시간 배양하였다. 이 때, 세포수는 5×104세포/플라스크였다. 여기에 적당 농도(10㎕, 50㎕)의 어성초 뿌리 추출물과 알부틴(동경화성제) 및 코직산(시그마제)을 DMEM 배지에 희석시켜 배양된 멜라노마 세포에 처리하여 37℃에서 5일간 배양하였다. 배양 후 배지를 모두 제거하고 0.02% EDTA와 0.05%트립신을 함유한 살린포스페이트 완충액(Phosphate buffered saline)을 1㎖ 처리하여 세포를 분리시킨, 후 5분간 원심분리(1500rpm)하여 침전된 멜라닌 생성량을 475㎚에서 흡광광도계(160-A, Shimazu제)로 측정하였다. 티로시나제 활성저해작용(%)은 시료미첨가시의 멜라닌 양에 대한 비율로서 구하였다.Inhibition of melanin biosynthesis of B16 melanoma cells against the fish extract and arbutin and kojic acid obtained in Example 1 was tested. Mouse-derived B16 melanoma cells (American Type Culture Collection, Accession No.:6323) were inoculated into DMEM (Dulbecco's Modified Eagles Medium (Gibco, USA)) medium containing 4.5 g / L glucose and 10% fetal bovine serum 1% antibiotic. Incubated at 37 ° C. in a 50 ml T-flask. After 24 hours of incubation under 5% CO 2 conditions, the cells were isolated by treatment with 0.05% trypsin containing 0.02% EDTA, and then incubated in a 50 ml T-flask for 48 hours. At this time, the cell number was 5 x 10 4 cells / flask. Here, Echocho root extract, arbutin (curing agent) and kojic acid (Sigma) of appropriate concentrations (10 μl, 50 μl) were diluted in DMEM medium, treated with cultured melanoma cells, and cultured at 37 ° C. for 5 days. After incubation, all of the medium was removed and the cells were separated by treatment with 1 ml of phosphinate buffered saline containing 0.02% EDTA and 0.05% trypsin. It measured by the absorbance photometer (160-A, the Shimazu make) at nm. The tyrosinase inhibitory activity (%) was calculated as a ratio with respect to the melanin amount when no sample was added.
상기 표에 나타난 바와 같이, 어성초추출물이 알부틴, 코직산에 비해 훨씬 우수한 미백효과를 보였다.As shown in the table, Echochocho extract showed much better whitening effect than arbutin and kojic acid.
시험예 3Test Example 3
인체사용시험Human Use Test
본 발명의 실시예 1∼5에서 얻은 어성초추출물 5 중량%를 함유한 크림상의 시료를 성인남녀 50명(남자25명, 여자 25명)을 대상으로 상박부내측 연한 피부에 부착하였다. 24시간 후에 피부의 자극, 홍반, 가려움, 알러지 등의 피부상태를 관찰하고 다시 24시간 후에 한번 더 관찰하였다.A creamy sample containing 5% by weight of the fish paste extracted in Examples 1 to 5 of the present invention was attached to the inner skin of the upper arm of 50 males and females (25 males and 25 females). After 24 hours, skin irritation, erythema, itching, and allergic skin conditions were observed and again 24 hours later.
상기 표 3에 나타난 바와 같이, 어성초추출물 5%(중량)를 함유한 크림은 실시예 2와 실시예 5의 어성초 뿌리추출물 함유 크림에서만 1명씩 양성반응을 보일 뿐 나머지 실시예에서는 양성반응을 보이지 않아, 인체피부에 대한 안전성이 우수하다는 것을 확인하게 되었다.As shown in Table 3, the cream containing 5% (by weight) of Echochocho extract only shows positive reactions by one person in the cream containing Echochocho root extracts of Examples 2 and 5, but did not show a positive response in the other examples. It was confirmed that the safety of human skin is excellent.
시험예 4Test Example 4
본 발명 화장료의 미백효과를 실제 사용 테스트를 통하여 평가하였다. 어성초 추출물을 함유한 제조예 4를, 어성초추출물을 함유하지 않은 것을 제외하고는 동일한 제법의 제조예 4를 대조군으로 하여, 안면에 기미만 발생하고 다른 피부질환이 없는 20 - 40세의 여성 20명에 대해서 기미 치유효과를 실험하였다.The whitening effect of the cosmetic of the present invention was evaluated through a practical use test. Twenty females of 20-40 years of age who did not have other skin diseases and had only spots on their face were prepared using Preparation Example 4, which contains Eochocho extract, and Preparation Example 4 of the same method, except that Echochocho extract did not contain Eochocho extract. The blemish healing effect was tested.
시험대상자 20명에게 안면의 왼쪽부위에는 어성초추출물을 함유한 제조예 4를, 오른쪽 부위에는 추출물을 함유하지 않은 제조예 4(대조군)를 매일 아침, 저녁 2회씩 세안 후 적당량을 2개월간 바르게 하였다. 그 결과를 육안으로 관찰하여 기미 부위의 탈색정도를 대조군과 비교하여 평가하였다.Twenty subjects were treated with Preparation Example 4, which contains Echo extract on the left side of the face, and Preparation Example 4 (Control), which did not contain the extract on the right side, after washing two times each morning and evening. The results were visually observed to evaluate the degree of discoloration of the spots by comparison with the control group.
상기 표 4에서 나타난 바와 같이, 어성초추출물을 함유한 본 발명의 화장품은 대조군에 비하여 훨씬 우수한 미백효과가 나타남을 알 수 있다.As shown in Table 4, it can be seen that the cosmetic of the present invention containing Echochocho extract has a much better whitening effect than the control.
본 발명에 의한 어성초추출물을 기존에 개발된 알부틴, 코직산과 비교한 결과, 어성초 추출물이 가장 뛰어난 티로시나제 활성 저해효과를 보임으로써 미백효과 뿐 만 아니라 안전성 면에서도 매우 우수함이 증명되었다.Compared with the arbutin and kojic acid, the extract of Echochocho extract according to the present invention was proved to be excellent in terms of safety as well as whitening effect by showing the best inhibitory tyrosinase activity.
또한, 상술한 어성초추출물은 피부 미백화장료에 적용이 가능하고 화장료에 배합시 우수한 미백효과가 나타났다.In addition, the above-mentioned fish vinegar extract can be applied to skin whitening cosmetics and showed an excellent whitening effect when blended in cosmetics.
Claims (6)
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100848515B1 (en) * | 2008-03-17 | 2008-07-25 | 주식회사 참선진 종합식품 | Cosmetics for whitening compring mushroom fermentation as an effective ingredient |
| KR101103283B1 (en) * | 2009-04-16 | 2012-01-11 | (주)휴럼 | The Manufacturing Method of Fermented Aloe for Whitening Effect in skin, and the Functional Whitening Cosmetics Containing Fermented Aloe |
| TWI480062B (en) * | 2012-05-23 | 2015-04-11 | Chu Biology Tech Co Ltd De | A use of an herbal extract of manufacturing preparations for skin whitening |
| KR20180013541A (en) * | 2016-07-29 | 2018-02-07 | 김윤 | liposomes encapsulating natural plant extracts and the method of manufacturing for preparing the same |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0717846A (en) * | 1993-06-30 | 1995-01-20 | Sansho Seiyaku Co Ltd | Skin external preparation |
| JPH10251155A (en) * | 1997-03-10 | 1998-09-22 | Shiseido Co Ltd | Protease inhibitor |
| KR20000058332A (en) * | 2000-04-21 | 2000-10-05 | 이성낙 | Cosmetic or pharmaceutical composition comprising Houttuynia cordata extract for preventing and curing acne |
| KR20000064232A (en) * | 2000-08-30 | 2000-11-06 | 유연실 | The massage make-up of wrinkles remove |
| KR20010044078A (en) * | 2000-08-19 | 2001-06-05 | 김완찬 | Manufacturing method of beauty cleansing water |
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- 2000-12-05 KR KR1020000073163A patent/KR20020044194A/en not_active Application Discontinuation
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0717846A (en) * | 1993-06-30 | 1995-01-20 | Sansho Seiyaku Co Ltd | Skin external preparation |
| JPH10251155A (en) * | 1997-03-10 | 1998-09-22 | Shiseido Co Ltd | Protease inhibitor |
| KR20000058332A (en) * | 2000-04-21 | 2000-10-05 | 이성낙 | Cosmetic or pharmaceutical composition comprising Houttuynia cordata extract for preventing and curing acne |
| KR20010044078A (en) * | 2000-08-19 | 2001-06-05 | 김완찬 | Manufacturing method of beauty cleansing water |
| KR20000064232A (en) * | 2000-08-30 | 2000-11-06 | 유연실 | The massage make-up of wrinkles remove |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100848515B1 (en) * | 2008-03-17 | 2008-07-25 | 주식회사 참선진 종합식품 | Cosmetics for whitening compring mushroom fermentation as an effective ingredient |
| KR101103283B1 (en) * | 2009-04-16 | 2012-01-11 | (주)휴럼 | The Manufacturing Method of Fermented Aloe for Whitening Effect in skin, and the Functional Whitening Cosmetics Containing Fermented Aloe |
| TWI480062B (en) * | 2012-05-23 | 2015-04-11 | Chu Biology Tech Co Ltd De | A use of an herbal extract of manufacturing preparations for skin whitening |
| KR20180013541A (en) * | 2016-07-29 | 2018-02-07 | 김윤 | liposomes encapsulating natural plant extracts and the method of manufacturing for preparing the same |
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